The NYSGRC has a 10-year proven track record in high-throughput structure determination as well as in discovering and implementing infrastructure to increase the speed, accuracy, success rate and affordability of structural biology studies. Moving forward the NYSGRC will reorganize to meet the new and diverse challenges associated with the PSI:Biology Network. In addition to reliance on traditional high-throughput bacterial expression platforms, the NYSGRC will develop and implement cutting-edge experimental and computational technologies to examine the biologically important molecules that are the focus of PSI:Biology. These targets are likely to include multidomain eukaryotic proteins, multi-component assemblies, and secreted proteins that underlay complex multi-cellular biology and directly contribute to human health and disease. In addition to servicing the High-Throughput-Enabled Structural Biology Partnerships, this infrastructure will support our Biological Theme that focuses on the secretion machinery and secreted effector proteins from major bacterial, protozoan and fungal pathogens. These targets were specifically selected to provide new insights into the mechanisms that these pathogens have evolved for immune evasion and modulation of host signaling pathways. These processes rely on intricate nano-machines, with cytoplasmic, membrane-associated and extracellular components, that require hybrid computational and experimental approaches to define their organization, structure and function. Finally, our integrated experimental and computational efforts have identified new opportunities to significantly and economically enhance sequence/structure coverage. The advent of PSI:Biology is driving a process of evolutionary change for the NYSGRC that has already enhanced its outstanding high-throughput structure determination pipeline. Our strengths in traditional bacterial expression, coupled with novel approaches to eukaryotic expression and refolding, as well as our established expertise in hybrid methods, positions us to uniquely support the efforts of PSI:Biology. PUBLIC HEALTH RELEVANCE: The NIH is committed to advancing research directly relevant to a detailed understanding of human health and disease states. The research proposed in this application is designed to significantly enhance the ability of the scientific community to define the shapes and structures of important macromolecules. This information provides enormous insights into strategies to develop drugs and therapeutics to combat infectious diseases, autoimmune diseases and cancers.